LSU VETERINARY COBRE: PATHOGENESIS OF RICKETTSIA SP

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 明确的目标。 目的1.描述SFG立克次体在单个硬体和单个硬质组织中的传播速度。未受感染的变色尘螨将通过毛细管喂养暴露于立克次体、蒙丹氏立克次体或贝氏立克次体。将确定SFG感染和传播的参数，包括分散和繁殖，在单个硬蜱组织中。 使用实时定量聚合酶链式反应评估立克次体感染和随后的组织传播。 在感染的卵、幼虫、若虫和成虫阶段，量化与水平(唾液腺)和垂直(卵巢)传播事件相关的立克次体。 目的2.鉴定和鉴定三种硬蜱携带的立克次体在水平和垂直传播事件中在硬体宿主中唯一表达的蛋白质。虽然还不是很清楚，但已经描述了其他SFG立克次体和它们的壁虱宿主之间的代谢结合。立克次体经历了细胞结构和脊椎动物感染性的重新激活事件，这与宿主环境线索相关联，与获取血粉有关。这一目标将提供有关壁虱传播的立克次体的全球蛋白表达和抗原谱的新信息。 答：最初的研究将通过双向凝胶电泳法比较SFG立克次体在水平(唾液腺)和垂直(卵巢)传播所涉及的扁虱组织中的传播事件中的蛋白质谱。 B.通过免疫印迹和四倍飞行时间(2DLC-QTOF)分析鉴定立克次体抗原。 C.将观察到的蛋白质表达与实时定量聚合酶链式反应评估的基因表达进行比较。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Specific Aims. Aim 1. Characterize the rate at which SFG rickettsiae spread through individual ticks and throughout individual tick tissues. Uninfected Dermacentor variabilis will be exposed to Rickettsia rickettsii, R. montanensis, or R. bellii via capillary feeding. Parameters of SFG infection and transmission, including dispersion and multiplication, in individual tick tissues will be determined. A. Assess SFG rickettsial infection and subsequent tissue dissemination in the tick vector using quantitative real-time PCR. B. Quantify rickettsiae associated with horizontal (salivary gland) and vertical (ovary) transmission events in infected egg, larva, nymph, and adult stages. Aim 2. Identify and characterize uniquely expressed protein from three tick-borne rickettsiae in tick hosts during horizontal and vertical transmission events. Although not well understood, a metabolic union between other SFG rickettsiae and their tick hosts has been described. Rickettsia undergo a reactivation event changing in cell structure and vertebrate infectivity in association with host environmental cues coupled to bloodmeal acquisition. This aim will provide novel information concerning tick-borne rickettsiae in relation to global protein expression and antigenic profiles. A. Initial studies will compare protein profiles of SFG rickettsiae during transmission events within the tick tissues involved in horizontal (salivary gland) and vertical (ovary) transmission by two dimensional gel electrophoresis. B. Identify rickettsial antigens by immunoblot using polyclonal antibodies to tick-borne rickettsiae and liquid chromatography quadruple-time-of-flight (2DLC-QTOF) analysis. C. Compare observed protein expression to gene expression, as assessed by real-time PCR.